Yao Yupeng, Lin Weifan, Zhang Yan
Key Laboratory of Gene Engineering of the Ministry of Education, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University.
Key Laboratory of Gene Engineering of the Ministry of Education, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University;
J Vis Exp. 2018 Jun 20(136):57235. doi: 10.3791/57235.
In order to construct an effective and realistic model for tongue squamous cell carcinoma (TSCC) in vitro, the methods were created to produce decellularized tongue extracellular matrix (TEM) which provides functional scaffolds for TSCC construction. TEM provides an in vitro niche for cell growth, differentiation, and cell migration. The microstructures of native extracellular matrix (ECM) and biochemical compositions retained in the decellularized matrix provide tissue-specific niches for anchoring cells. The fabrication of TEM can be realized by deoxyribonuclease (DNase) digestion accompanied with a serious of organic or inorganic pretreatment. This protocol is easy to operate and ensures high efficiency for the decellularization. The TEM showed favorable cytocompatibility for TSCC cells under static or stirred culture conditions, which enables the construction of the TSCC model. A self-made bioreactor was also used for the persistent stirred condition for cell culture. Reconstructed TSCC using TEM showed the characteristics and properties resembling clinical TSCC histopathology, suggesting the potential in TSCC research.
为了构建一种有效的、符合实际的体外舌鳞状细胞癌（TSCC）模型，人们创建了制备脱细胞舌细胞外基质（TEM）的方法，该基质为TSCC模型构建提供功能性支架。TEM为细胞生长、分化和细胞迁移提供体外微环境。天然细胞外基质（ECM）的微观结构以及保留在脱细胞基质中的生化成分，为锚定细胞提供了组织特异性微环境。TEM的制备可通过脱氧核糖核酸酶（DNase）消化并辅以一系列有机或无机预处理来实现。该方案操作简便，可确保脱细胞过程高效进行。在静态或搅拌培养条件下TEM对TSCC细胞显示出良好的细胞相容性，这使得TSCC模型的构建成为可能。还使用了自制的生物反应器进行持续搅拌细胞培养。使用TEM重建的TSCC表现出类似于临床TSCC组织病理学的特征和特性，表明其在TSCC研究中的潜力。
